The purpose of this proposal is a detailed study of the immunoreactivity of various isocyanate protein conjugates in several experimental animal models. In earlier work, our laboratory clearly demonstrated that hapten-specific murine homocytotropic antibodies could be produced after parenteral immunization. In these studies, it was also established that proper chemical characterization and stability of immunizing reagents was essential. The current studies will continue to develop better methods of preparing and characterizing various monomeric and polymeric compounds which are formed during the reaction of isocyanates with proteins carrier. In this extended work immunogens will consist of monomeric isocyanate protein conjugates. The ability of these substances to induce homocytotropic antibody and IgG immune responses in mice, rabbits and guinea pigs will be studied by standard immunologic methods. Induction of cell mediated immunity by these substances will also be studied by delayed type skin testing and the macrophage inhibitory factor test in guinea pigs. In addition, induction of contact dermatitis by various isocyanate compounds, applied directly to guinea pigs' skin will also be attempted. In other experiments the effects of chronic exposure to isocyanates will be monitored in guinea pigs by immunologic reactivity, serial pulmonary physiologic changes, the possible development of anaphylactic sensitivity and pathologic changes at the conclusion of the experiment. Monoclonal mouse IgG and IgE-antibodies with isocyanate specificity will also be produced by preparing secreting hybridomas. Larger amounts of antibody obtained by these methods will permit other experiments dealing with antigen induced histamine release by passively sensitized mast cells, regulatory factors of antibody synthesis and the preparation of fluorescent antibodies for direct examination of antigen localization within tissues.